Structural studies will be performed on a series of H3-labeled oligosaccharides obtained by hydrolytic or deaminative cleavage of H3- carboxyl-reduced heparin and heparan sulfate. Several unidentified monosaccharides derived from heparin or carboxyl-reduced heparin will be characterized by mass spectrometry. As a further aid in the determination of the heparin structure attempts will be made to develop improved procedures for sulfate analysis. The procedures which have been developed in this laboratory for radio-chemical determination of glycogen, mucopolysaccharides, glycolipids, sialic acid, and metabolic pools will be used in the study of the carbohydrates of normal and virally transformed chick embryo fibroblasts, as well as various chick embryo tissues and cultured cells. Further procedures for analysis of heparin and heparan sulfate in chick embryos will be developed. The primary tool that will be used will be a purified Flavobacterium heparinum heparinase obtained by affinity chromatography on columns prepared by coupling carboxyl-reduced heparin to agarose. Finally mapping procedures will be developed for comparing cell surface glycoproteins or glycopeptides obtained from different embryonic tissues.